The effect of calcium phosphate nanoparticles⁃mediated interference of LMO4 on skin squamous cells / 安徽医科大学学报

ABSTRACT

Objective @#To investigate lim domain protein 4 (LMO4) functions and mechanisms in regulating proliferation of skin squamous cells (A431) , the shRNAs targeted to human LMO4 were coated by calcium phosphate nanoparticles (NP) and transfected into A431 cells to inhibit LMO4 expression. @*Methods @#Reverse transcription and quantitative polymerase chain reaction ( RT⁃qPCR) , immunohistochemistry analysis and Western blot were used to detect expression of the interest genes. The expression vectors with shRNA targeted to human LMO4 (NP/sh⁃L) were coated by the calcium phosphate nanoparticles , and transfected into A431 . The MTT assay was conducted to determine cell proliferation after transfected for 24 , 36 and 48 h. Cells were stained with propidium iodide and examined cell cycles by using flow cytometry. @*Results @# LMO4 expressed at higher levels both in the skin squacalcium phosphate nanoparticles and DNA was 10 ∶ 1 . There was no significant difference of transfection efficiency between the NP/sh⁃L and lipofection approaches. The MTT assay showed that silencing LMO4 inhibited proliferadown did not alter expression of CDK4 and cyclin D1 .@*Conclusion@#The calcium phosphate nanoparticles could bind and transfer the foreign DNA into the targeted cells with high efficiency. Silencing LMO4 decreased expression of cyclin E and CDK2 resulted in inhibition of cell proliferation.